Instrumental Methods of Analysis

Analytical chemistry deals with the quantitative and qualitative characterization of materials.
The identification of elements/ radicals/ ions/ compounds is called as qualitative analysis.
The determination of exact quantity of the constituents present in the substance is called as
quantitative analysis. A chemical analysis which involves instruments is called as
instrumental analysis.
Advantages
1. It is faster.
2. We get accurate and reproducible result.
3. Low concentration sample solution can be analyzed.
4. Easy to handle large number of samples.
5. Widely applicable in industries.

Disadvantages
1. Instruments are expensive.
2. Calibration is necessary before analysis.
3. Concentration range is limited.
4. Training is needed to handle the instruments.

Potentiometry
Potentiometry is an instrumental analysis in which the amount of a substance in solution is
determined by measuring the EMF between two electrodes dipped in the solution.
The potential of an electrode is given by Nernst equation.
E = E0 + (0.0591/n) log [Mn+]
Thus, the potential of an electrode depends upon the concentration of the ion.
Electrode potential is measured by combining the electrode with reference electrode. During
titration, concentration of ions changes. Hence potential of the electrode changes. The
electrode, whose potential depends on the concentration of the ions to be determined, is
called as indicator electrode.
Instrumentation
The potentiometer consists of a reference electrode, an indicator electrode and a potential
measuring device. A simple arrangement is as shown in the figure.

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A know volume of analyte is taken in a beaker and its potential is determined. The titrant is
added in increment of 0.5cm3 and emf is measured for each addition. At the approach of the
equivalent point the emf tends to increase rapidly. A few reading are taken beyond the end
point. Thus the change in potential at different volume of titrant are recorded. To locate the
end point of the titration emf value are plotted against the volume of titrant or ∆E/∆V values
are plotted against the volume of titrant.

Applications
1. To locate end point of the titration.
2. To determine the concentration of the solution by finding emf.
3. To determine pH of the solution using glass electrode.

Potentiometric titration is applicable in oxidation-reduction titration to estimate the

concentration of analyte.
In redox titration the indicator electrode is an inert metal such as platinum and the reference
electrode is generally a saturated calomel electrode.
The potential of the electrode depends on the concentration of the substance being oxidized
or reduced.
E = Eo + (0.0591/n) log {[oxidized] / [reduced]}
E = Eo + (0.0591/n) log {[Fe3+] / [Fe2+]}
Immerse a platinum electrode and a reference electrode (calomel electrode) into the given
FAS solution in the beaker and connect it to the potentiometer. Add one test tube of dilute
H2SO4 solution. Measure the emf of the solution. Add standard solution of K 2CrO7 from the
burette in increment of 0.5 cm3. After each addition stir the solution and note the emf.
Continue the titration till a sharp increase in emf is obtained and 5 to 6 reading thereafter.
Plot ∆E/∆V against volume of K2Cr2O7 and note down the equivalence point.

Colorimetry
Colorimetry is an instrumental analysis in which the amount of a substance in solution which
are themselves coloured or which gives colour with a suitable reagent is determined.
Colorimetric method of analysis is based on Beer’s and Lambert’s Law.
Beer’s law - states that the intensity of the transmitted light decreases exponentially as the
concentration of the media increases arithmetically.
Lambert’s law- states that the intensity of the transmitted light decreases exponentially as
the path length of the media increases arithmetically.

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 Instrumental Methods of Analysis
Beer’s Lambert’s law states that when a monochromatic light is passed through a solution,
part of light is absorbed by the solution. The extent of absorption depends on the
concentration of the solution and the path length of the light through the solution.
A = εcl
A = absorbance c = concentration of the solution, l = path length ε = molar absorption
coefficient.
If the path length l of the cell is kept constant, absorbance A is proportional to the
concentration C.
AαC
Therefore if absorbance of a series of solution are plotted versus their concentration, a
straight line graph result, which is called Beer’s-Lambert’s curve or calibration curve.

Instrumentation
The instrument used to measure the absorbance of a solution is called photoelectric
colorimeter. It consists of,
1. Tungsten lamp as light source
2. A filter which provides the desired wavelength range, where in the solution gives the
maximum absorbance.
3. A sample cell
4. A photocell detector

Blank solution is taken in the sample cell and placed in the path of the light beam. Its
absorbance is adjusted to zero on the meter. Light from tungsten lamp is allowed to fall on
the solution after passing through the filter. The quantity of light absorbed is measured. A plot
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of absorbance versus concentration of analyte is used to find the unknown concentration of
analyte in the sample.
Applications
1. Used in chemical lab for the analysis of glucose, urea creatinine etc
2. Used in soil testing lab
3. Used in water and food testing lab

Example: Colorimetry is used in the estimation of copper in a solution. Cupric ions react with
ammonia to give a deep blue colour due to the formation of cuprammonium complex ion.
Cu2+ + 4NH3  [Cu(NH3)4]2+
Procedure
Prepare 100ml of 0.1M solution of CuSO4. Transfer 5, 10, 15, 20, 25 ml of prepared CuSO 4
solution into different 50 ml volumetric flasks using burette. Add 5 ml of ammonia solution to
all the flasks and make up to the mark using distilled water. Stopper the flask and shake
well.
Prepare a blank solution by taking 5 ml of NH3 solution in 50 ml of volumetric flask, make up
to the mark by using distilled water and shake well. Measure absorbance of all the solutions
against blank at 620 nm using a photoelectric colorimeter.
To the given unknown solution add 5 ml of NH 3 and make up to the mark with distilled water.
Then a calibration curve is obtained by plotting absorbance against concentration. From the
graph the concentration of copper ions is determined.

Conductometry
Conductometry is an instrumental analysis in which the amount of a substance in solution
can be determined by measuring conductance of the solution.
Theory: Conductivity of the electrolyte solution obeys Ohm’s law and at constant temperature
it depends on charge, mobility, concentration and size of the ions. Ohm’s law states that the
current I flowing through a conductor is directly proportional to the applied potential E and
inversely proportional to the resistance R of the conductor.
E = IR
Instrumentation

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It consists of two platinum electrodes each of unit area of cross section placed unit distance
apart. The electrodes are dipped in the electrolyte solution taken in a beaker. Two electrodes
are connected to a conductometer and the conductance of the solution is recorded. The
titrant is added from the burette in an increment of 0.5 ml. The conductance is measured
after each addition. Then a graph of conductance against the volume of titrant is plotted. The
point of intersection of the curve gives the end point.

Application
1. Strong acid with strong base (HCl v/s NaOH)
NaOH + HCl  NaCl+ H2O
A graph of conductance against the volume of NaOH is as shown in the figure. On addition of
NaOH, the conductance decreases because the small and highly mobile H + ions in the acid
is replaced by heavier and less mobile Na+ ions. Addition of base beyond the neutralization
point leads to increase in the more mobile OH- ions and conductance increases.

2. Weak acid and strong base (CH3COOH v/s NaOH)

NaOH + CH3COOH  CH3COONa + H2O
A graph of conductance against the volume of NaOH is as shown in the figure. On addition of
NaOH, the conductance increases because the partially dissociated weakly conducting weak
acid is converted into fully dissociated conducting salt (sodium acetate). Addition of base
beyond the neutralization point leads to increase in the more mobile OH - ions and
conductance increases.

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3. Acid mixture with strong base (HCl and CH3COOH v/s NaOH)
A graph of conductance against the volume of NaOH is as shown in the figure. On addition of
NaOH, the conductance decreases because the small and highly mobile H + ions in the acid
is replaced by heavier and less mobile Na+ ions.
NaOH + HCl  NaCl+ H2O
addition of base beyond the first neutralization point leads to moderate increase in
conductance because the partially dissociated poorly conducting weak acid is converted into
fully dissociated conducting salt (sodium acetate) until the second neutralization point.

Further addition of base increases the number of more mobile OH - ions and hence
conductance increases.

Flame Photometry (Flame Emission Spectroscopy)

Flame Photometry is an instrumental analysis in which alkali and alkaline earth metals such
as Na, K, Li, Ca, Ba….are detected and determined. These elements emits characteristic
colour to Bunsen flame and the brightness of the flame is directly proportional to the
concentration of the element.
Following changes takes place when a solution containing a compound of the metal to be
investigated is aspirated into a flame.
1. Solvent evaporates leaving behind a solid residue in the form of mist.
2. Vaporization of the solid followed by its dissociation into constituent atoms which are
initially in the ground state.
3. Some gaseous atoms get excited by thermal energy of the flame from the ground state to
higher energy level (exited state).
4. Excited atoms are unstable and quickly emit photons and return to lower energy state’
5. The intensity of the emitted light is measured using flame photometer.

M+X-(aq) flame M+X-(Mist) evaporation MX (s)vaporization MX (g) Dissociation M(g) + X (g)

Thermal excitation

M* (g) excited atom

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The characteristic wavelength of some metal are Na-589 nm, Ca-623 nm, Li-670 nm, K-766
nm, Ba-554 nm.
Instrumentation

A simple flame photometer consists of the following basic components:

1. Source of flame
2. Nebulizer
3. Optical system
4. Simple colour filters
5. Photo-detector
Applications
 Flame photometer can be applied both for quantitative and qualitative analysis of elements. The
radiations emitted by the flame photometer are characteristic to particular metal. Hence with the
help of Flame photometer we can detect the presence of any specific element in the given sample.
 The presence of some group II elements is critical for soil health. We can determine the presence
of various alkali and alkaline earth metals in soil sample by conducting flame test and then the soil
can be supplied with specific fertiliser.
 The concentrations of Na+ and K+ ions are very important in the human body for conducting
various metabolic functions. Their concentrations can be determined by diluting and aspirating
blood serum sample into the flame.
 Soft drinks, fruit juices and alcoholic beverages can also be analysed by using flame photometry to
determine the concentrations of various metals and elements.
Example: Determination of Sodium
Prepare 1 ppm of NaCl solution. Transfer 2, 4, 6, 8 and 10ml of standard NaCl solution into
25ml of volumetric flask and dilute up to the mark with distilled water. Measure the emission
intensity of each solution and test solution using flame photometry.
Draw a calibration curve by plotting emission intensity against the concentration of NaCl
solution. From the calibration curve, the concentration of sodium in test solution can be
obtained.

Atomic Absorption Spectroscopy (AAS)

When a solution containing the sample element is aspirated into the flame, solvent gets
evaporated leaving behind the salt in the flame. The salt gets evaporated into vapour of the
salt, which further undergo dissociation into the constituent atoms. These metal atoms
electrons are excited by the flame energy, but do not have the necessary energy to jump to
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next quantum energy level. If a light of appropriate energy (wave length) is passed through
the flame, the excited electrons absorbs this wavelength photons and jambs to next energy
level.
M+X-(aq) flame M+X-(Mist) evaporation MX (s)vaporization MX (g) Dissociation M(g) + X (g)

Absorption of radiation

M* (g) excited atom

The grater the concentration of metal atoms in solution then more photons are absorbed in
the flame. This follows Beer’s –Lambert’s law.
A = εcl
A = absorbance c = concentration of the solution l = path length ε = molar absorption
coefficient.
The signal strength after the flame, decreases with increase in concentration. So by
measuring the absorbance by a suitable detector, the concentration can be determined.
Instrumentation

Atomic absorption spectrometer consists of

1. Hollow cathode lamp: is a source of light in which cathode is coated with a metal to be
analysed.
2. Nebulizer: It produces fine spray of the sample and introduce it into the flame.
3. Burner: here the sample is burned with the flame produced by the fuel and oxidant. The
sample after evaporation and vapourization leaves a residue of neutral atom.
4. Fuel and oxidant: proper combination of fuel and oxidant are used to produce required
temperature commonly used fuel are propane, hydrogen and acetylene and oxidants are air
or oxygen.
5. Monochromator and detector: the radiation from the hallow cathode lamp is passed
through a monochromator to separate the element specific radiation from other radiation
emitted by the radiation source and finally measured by a detector.
Applications
1. Used in the quantitative analysis of metal element in soil sample, body fluids,
environmental sample like ground water.
2. Used in food industries.
3. Estimate lead in petroleum product.

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